School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China.
School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China; Research Laboratory of Agricultural Environment and Food Safety, Anhui Modern Agricultural Industry Technology System, Hefei 230009, China.
Int J Biol Macromol. 2020 Nov 1;162:1303-1310. doi: 10.1016/j.ijbiomac.2020.06.246. Epub 2020 Jun 27.
Development of nontoxic, targetable and potent small interfering RNAs (siRNA) delivery systems remains a predominant challenge for clinical application of siRNA therapy. The nanoparticles of carboxymethyl chitosan (CMC) and labeled fluorescein isothiocyanate (FITC)-chitosan hydrochloride (CHC) were fabricated as carriers for ultrasound-triggered drug delivery to treat colon cancer. The results showed the (FITC-CHC)-CMC nanoparticles could effectively encapsulate anti-β-catenin siRNA through ionic gelation self-assembly to improve the stability of siRNA. The cumulative release ratio of siRNA from crosslinked (FITC-CHC)-CMC nanoparticles was merely 11.08% in pH 2.2 solution within 120 min, whereas about 70.07% of the loaded siRNA was released within 120 min in pH 5.5 solution after an 8-min ultrasonic treatment. It indicated that the (FITC-CHC)-CMC based pH-sensitive delivery system could fulfill a controlled release of siRNA through responding to external stimulus (ultrasound) under favorable pH condition. Fluorescence microscopy measurements clearly visualized the entry of fluorescently-labeled siRNA into HT-29 cells. Following the transfection of anti-β-catenin siRNA for 48 h, the β-catenin protein expression of the colon cancer cells was reduced to about 40.10%, indicating effective reduction of the protein that promotes colon cancer proliferation. Our results demonstrated that the siRNA-(FITC-CHC)-CMC delivery system hold substantial potential for RNAi therapeutical applications in diseased cells.
开发无毒、靶向和有效的小干扰 RNA（siRNA）递药系统仍然是 siRNA 治疗临床应用的主要挑战。羧甲基壳聚糖（CMC）纳米粒和标记有荧光素异硫氰酸酯（FITC）-壳聚糖盐酸盐（CHC）的纳米粒被制成载体，用于超声触发递药治疗结肠癌。结果表明，（FITC-CHC）-CMC 纳米粒可以通过离子凝胶自组装有效包载抗-β-catenin siRNA，提高 siRNA 的稳定性。交联的（FITC-CHC）-CMC 纳米粒在 pH 2.2 溶液中 120 min 内的 siRNA 累积释放率仅为 11.08%，而在 pH 5.5 溶液中经 8 min 超声处理后，约 70.07%的负载 siRNA 在 120 min 内释放。这表明基于（FITC-CHC）-CMC 的 pH 敏感递药系统可以通过在有利的 pH 条件下对外界刺激（超声）作出响应来实现 siRNA 的控制释放。荧光显微镜测量清楚地显示了荧光标记的 siRNA 进入 HT-29 细胞。转染抗-β-catenin siRNA 48 h 后，结肠癌细胞的β-catenin 蛋白表达降低到约 40.10%，表明促进结肠癌增殖的蛋白有效减少。我们的结果表明，siRNA-（FITC-CHC）-CMC 递药系统在病变细胞中具有很大的 RNAi 治疗应用潜力。
